LOCUS       sequence-311907-       10825 bp    DNA     circular SYN 14-APR-2025
DEFINITION  A multicistronic vector with CAGGS promoter-driven AsCpf1 and U6 
            promoter-driven single guide RNA (sgRNA) targeting the human INS 
            locus (sgINS)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10825)
  AUTHORS   Ma H, Jeppesen JF, Jaenisch R
  TITLE     Human T Cells Expressing a CD19 CAR-T Receptor Provide Insights into
            Mechanisms of Human CD19-Positive beta Cell Destruction.
  JOURNAL   Cell Rep Med. 2020 Sep 22;1(6):100097. doi: 
            10.1016/j.xcrm.2020.100097. eCollection 2020 Sep 22.
  PUBMED    33205073
REFERENCE   2  (bases 1 to 10825)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep 
            Med."; date: "2020-09-22"; volume: "1(6):100097. doi"; pages: " 
            10.1016/j.xcrm.2020.100097. eCollection 2020 Sep 22"
COMMENT     Sequence Label: CAGGS-AsCpf1-2A-GFP-U6-INS-sgRNA
FEATURES             Location/Qualifiers
     source          1..10825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        62..365
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..644
                     /label=chicken beta-actin promoter
     intron          645..1661
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1585..1607
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1669..1688
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1728..1748
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1773..5690
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
                     YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
                     IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
                     FTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
                     NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN
                     LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
                     NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
                     ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
                     KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
                     NYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     regulatory      5410..5419
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5691..5738
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5745..5825
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             5835..5888
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(5889..5891,5892..6602)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5889..5891,5892..6602)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5931..5952)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6192..6211)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6539..6560
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6630..6647)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6636..6843
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6852..6992
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6852..6981
                     /label=AAV2 ITR
     rep_origin      7067..7522
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7154..7173)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7364..7385
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7539..7558)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7658..7680
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7718..7736)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7804..7908
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7909..7977,7978..8769)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8127..8146)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8940..9528
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9429..9448
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        9590..9838
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     9590..9610
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9761..9780
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9889..9964
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     10188..10207
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    10328..10789
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
ORIGIN
        1 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
       61 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      121 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      181 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      241 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      301 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      361 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
      421 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
      481 ggggggggcg cgcgccgggg gggggggggg ggggsggggg gsggggsggg gcgaggcgga
      541 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
      601 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
      661 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
      721 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
      781 gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc
      841 gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg
      901 ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg
      961 ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
     1021 gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag
     1081 cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg
     1141 ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg
     1201 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg
     1261 gggrgggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg
     1321 cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt
     1381 cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg
     1441 gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc
     1501 cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc
     1561 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
     1621 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     1681 ttattgtgct gtctcatcat tttggcaaag accggtccac catggcccca aagaagaagc
     1741 ggaaggtcgg tatccacgga gtcccagcag ccacacagtt cgagggcttt accaacctgt
     1801 atcaggtgag caagacactg cggtttgagc tgatcccaca gggcaagacc ctgaagcaca
     1861 tccaggagca gggcttcatc gaggaggaca aggcccgcaa tgatcactac aaggagctga
     1921 agcccatcat cgatcggatc tacaagacct atgccgacca gtgcctgcag ctggtgcagc
     1981 tggattggga gaacctgagc gccgccatcg actcctatag aaaggagaaa accgaggaga
     2041 caaggaacgc cctgatcgag gagcaggcca catatcgcaa tgccatccac gactacttca
     2101 tcggccggac agacaacctg accgatgcca tcaataagag acacgccgag atctacaagg
     2161 gcctgttcaa ggccgagctg tttaatggca aggtgctgaa gcagctgggc accgtgacca
     2221 caaccgagca cgagaacgcc ctgctgcgga gcttcgacaa gtttacaacc tacttctccg
     2281 gcttttatga gaacaggaag aacgtgttca gcgccgagga tatcagcaca gccatcccac
     2341 accgcatcgt gcaggacaac ttccccaagt ttaaggagaa ttgtcacatc ttcacacgcc
     2401 tgatcaccgc cgtgcccagc ctgcgggagc actttgagaa cgtgaagaag gccatcggca
     2461 tcttcgtgag cacctccatc gaggaggtgt tttccttccc tttttataac cagctgctga
     2521 cacagaccca gatcgacctg tataaccagc tgctgggagg aatctctcgg gaggcaggca
     2581 ccgagaagat caagggcctg aacgaggtgc tgaatctggc catccagaag aatgatgaga
     2641 cagcccacat catcgcctcc ctgccacaca gattcatccc cctgtttaag cagatcctgt
     2701 ccgataggaa caccctgtct ttcatcctgg aggagtttaa gagcgacgag gaagtgatcc
     2761 agtccttctg caagtacaag acactgctga gaaacgagaa cgtgctggag acagccgagg
     2821 ccctgtttaa cgagctgaac agcatcgacc tgacacacat cttcatcagc cacaagaagc
     2881 tggagacaat cagcagcgcc ctgtgcgacc actgggatac actgaggaat gccctgtatg
     2941 agcggagaat ctccgagctg acaggcaaga tcaccaagtc tgccaaggag aaggtgcagc
     3001 gcagcctgaa gcacgaggat atcaacctgc aggagatcat ctctgccgca ggcaaggagc
     3061 tgagcgaggc cttcaagcag aaaaccagcg agatcctgtc ccacgcacac gccgccctgg
     3121 atcagccact gcctacaacc ctgaagaagc aggaggagaa ggagatcctg aagtctcagc
     3181 tggacagcct gctgggcctg taccacctgc tggactggtt tgccgtggat gagtccaacg
     3241 aggtggaccc cgagttctct gcccggctga ccggcatcaa gctggagatg gagccttctc
     3301 tgagcttcta caacaaggcc agaaattatg ccaccaagaa gccctactcc gtggagaagt
     3361 tcaagctgaa ctttcagatg cctacactgg cctctggctg ggacgtgaat aaggagaaga
     3421 acaatggcgc catcctgttt gtgaagaacg gcctgtacta tctgggcatc atgccaaagc
     3481 agaagggcag gtataaggcc ctgagcttcg agcccacaga gaaaaccagc gagggctttg
     3541 ataagatgta ctatgactac ttccctgatg ccgccaagat gatcccaaag tgcagcaccc
     3601 agctgaaggc cgtgacagcc cactttcaga cccacacaac ccccatcctg ctgtccaaca
     3661 atttcatcga gcctctggag atcacaaagg agatctacga cctgaacaat cctgagaagg
     3721 agccaaagaa gtttcagaca gcctacgcca agaaaaccgg cgaccagaag ggctacagag
     3781 aggccctgtg caagtggatc gacttcacaa gggattttct gtccaagtat accaagacaa
     3841 cctctatcga tctgtctagc ctgcggccat cctctcagta taaggacctg ggcgagtact
     3901 atgccgagct gaatcccctg ctgtaccaca tcagcttcca gagaatcgcc gagaaggaga
     3961 tcatggatgc cgtggagaca ggcaagctgt acctgttcca gatctataac aaggactttg
     4021 ccaagggcca ccacggcaag cctaatctgc acacactgta ttggaccggc ctgttttctc
     4081 cagagaacct ggccaagaca agcatcaagc tgaatggcca ggccgagctg ttctaccgcc
     4141 ctaagtccag gatgaagagg atggcacacc ggctgggaga gaagatgctg aacaagaagc
     4201 tgaaggatca gaaaacccca atccccgaca ccctgtacca ggagctgtac gactatgtga
     4261 atcacagact gtcccacgac ctgtctgatg aggccagggc cctgctgccc aacgtgatca
     4321 ccaaggaggt gtctcacgag atcatcaagg ataggcgctt taccagcgac aagttctttt
     4381 tccacgtgcc tatcacactg aactatcagg ccgccaattc cccatctaag ttcaaccaga
     4441 gggtgaatgc ctacctgaag gagcaccccg agacacctat catcggcatc gatcggggcg
     4501 agagaaacct gatctatatc acagtgatcg actccaccgg caagatcctg gagcagcgga
     4561 gcctgaacac catccagcag tttgattacc agaagaagct ggacaacagg gagaaggaga
     4621 gggtggcagc aaggcaggcc tggtctgtgg tgggcacaat caaggatctg aagcagggct
     4681 atctgagcca ggtcatccac gagatcgtgg acctgatgat ccactaccag gccgtggtgg
     4741 tgctggagaa cctgaatttc ggctttaaga gcaagaggac cggcatcgcc gagaaggccg
     4801 tgtaccagca gttcgagaag atgctgatcg ataagctgaa ttgcctggtg ctgaaggact
     4861 atccagcaga gaaagtggga ggcgtgctga acccatacca gctgacagac cagttcacct
     4921 cctttgccaa gatgggcacc cagtctggct tcctgtttta cgtgcctgcc ccatatacat
     4981 ctaagatcga tcccctgacc ggcttcgtgg accccttcgt gtggaaaacc atcaagaatc
     5041 acgagagccg caagcacttc ctggagggct tcgactttct gcactacgac gtgaaaaccg
     5101 gcgacttcat cctgcacttt aagatgaaca gaaatctgtc cttccagagg ggcctgcccg
     5161 gctttatgcc tgcatgggat atcgtgttcg agaagaacga gacacagttt gacgccaagg
     5221 gcaccccttt catcgccggc aagagaatcg tgccagtgat cgagaatcac agattcaccg
     5281 gcagataccg ggacctgtat cctgccaacg agctgatcgc cctgctggag gagaagggca
     5341 tcgtgttcag ggatggctcc aacatcctgc caaagctgct ggagaatgac gattctcacg
     5401 ccatcgacac catggtggcc ctgatccgca gcgtgctgca gatgcggaac tccaatgccg
     5461 ccacaggcga ggactatatc aacagccccg tgcgcgatct gaatggcgtg tgcttcgact
     5521 cccggtttca gaacccagag tggcccatgg acgccgatgc caatggcgcc taccacatcg
     5581 ccctgaaggg ccagctgctg ctgaatcacc tgaaggagag caaggatctg aagctgcaga
     5641 acggcatctc caatcaggac tggctggcct acatccagga gctgcgcaac aaaaggccgg
     5701 cggccacgaa aaaggccggc caggcaaaaa agaaaaaggg atcctaccca tacgatgttc
     5761 cagattacgc ttatccctac gacgtgcctg attatgcata cccatatgat gtccccgact
     5821 atgccggcag tggagagggc agaggaagtc tgctaacatg cggtgacgtc gaggagaatc
     5881 ctggcccagt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     5941 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     6001 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     6061 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     6121 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     6181 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     6241 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     6301 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     6361 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     6421 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     6481 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     6541 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     6601 aggaattcta actagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     6661 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     6721 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     6781 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gagaatagca ggcatgctgg
     6841 ggagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     6901 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     6961 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     7021 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     7081 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     7141 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     7201 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     7261 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     7321 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     7381 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     7441 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     7501 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     7561 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     7621 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     7681 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     7741 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     7801 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     7861 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     7921 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     7981 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     8041 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     8101 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     8161 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     8221 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     8281 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     8341 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     8401 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     8461 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     8521 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     8581 gctggctggt ttattgctga taaatctgga gccggtgagc gtggaagccg cggtatcatt
     8641 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     8701 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     8761 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     8821 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     8881 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     8941 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     9001 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     9061 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     9121 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     9181 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     9241 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     9301 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     9361 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     9421 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     9481 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     9541 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtg agggcctatt
     9601 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
     9661 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
     9721 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
     9781 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgt
     9841 aatttctact cttgtagatt tccatctctc tcggtgcagg agttttttgt tttagagcta
     9901 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     9961 gtgctttttt gttttagagc tagaaatagc aagttaaaat aaggctagtc cgtttttagc
    10021 gcgtgcgcca attctgcaga caaatggctc taggcatgcc gacggcgagg atctcgtcgt
    10081 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
    10141 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
    10201 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
    10261 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagg
    10321 ggatccgctg taagtctgca gaaattgatg atctattaaa caataaagat gtccactaaa
    10381 atggaagttt ttcctgtcat actttgttaa gaagggtgag aacagagtac ctacattttg
    10441 aatggaagga ttggagctac gggggtgggg gtggggtggg attagataaa tgcctgctct
    10501 ttactgaagg ctctttacta ttgctttatg ataatgtttc atagttggat atcataattt
    10561 aaacaagcaa aaccaaatta agggccagct cattcctccc actcatgatc tatagatcta
    10621 tagatctctc gtgggatcat tgtttttctc ttgattccca ctttgtggtt ctaagtactg
    10681 tggtttccaa atgtgtcagt ttcatagcct gaagaacgag atcagcagcc tctgttccac
    10741 atacacttca ttctcagtat tgttttgcca agttctaatt ccatcagaag ctgggagctc
    10801 caccgcggtg gcggccgctc tagaa
//
